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Establishing cell lines from fresh or cryopreserved tissue from the great crested newt (Triturus cristatus): a preliminary protocol

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  • Julie Strand, Aalborg University, Denmark
  • Henrik Callesen
  • Cino Pertoldi, Aalborg University, Institut for Kemi og Bioteknologi, Denmark
  • Stig Purup

This study describes a successful protocol for establishing cell lines from the threatened Triturus cristatus in terms of collection, preparing, establishing, cryopreserving, thawing and quality checking. Different parameters such as media, media change, fresh vs. cryopreserved tissue and seeding density were tested to optimize culture conditions for this species. With fresh tissue, no considerable differences in the use of two different media were found, but with cryopreserved tissue, a combination of ITS (insulin/transferrin/selenite) and 2-mercaptoethanol had a positive effect on growth. Real-time measurements on the cell lines were used, for the first time in amphibian cells, to investigate the effect of different treatments such as media change with or without washing. Media change had a positive impact on the cells, whereas the effect was negative when combined with washing. It is concluded that establishment of cell lines is possible from the great crested newt, especially when using fresh tissue, but much more challenging if the tissue has been cryopreserved. Real-time measurement during cell culture is a useful tool to visualize the sensitivity of amphibian cells during different culture treatments.

Original languageEnglish
Article number367
Number of pages12
Publication statusPublished - Feb 2021

    Research areas

  • Amphibian, Biobanking, Cell culture, Cryopreservation, Culture conditions, Morphology, XCELLigence®, real-time cell analysis

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